Beyond the Identification of Transcribed Sequences:
Functional, Evolutionary and Expression Analysis
12th International Workshop
October 25-28, 2002
Washington, DC

List of Abstracts * Speakers * Organizers * Authors * Original Announcement

Large-Scale Subcellular Localisation and Functional Analysis of Novel Proteins

Jeremy C. Simpson1, Urban I. Liebel1, Vytaute Starkuviene1, Holger Erfle1,  Annemarie Poustka2, Stefan Wiemann2 and Rainer Pepperkok1
1Cell Biology and Biophysics Programme, EMBL, 69117 Heidelberg, Germany and 2Molecular Genome Analysis, DKFZ, 69120 Heidelberg, Germany
Telephone: +49 6221 387232
Fax: +49 6221 387306

One of the greatest challenges facing biology today is the conversion of huge amounts of genomic data into more tangible functional information about the proteins encoded. For example, the large-scale cDNA sequencing project of the German cDNA Consortium is providing us with vast numbers of open reading frames (ORFs) encoding novel proteins of completely unknown function.As a first step towards their characterisation we have tagged over 300 of these with the green fluorescent protein (GFP), and examined the subcellular localisations of these fusion proteins in living cells. These data have allowed us to classify  these proteins into subcellular groups which in turn determines the next appropriate steps to characterise them. To make further use of these GFP-tagged constructs, a series of functional assays have been designed and implemented to assess the effect of these novel proteins on processes such as cell growth, organelle morphology and protein transport.

Functional assays with such a large set of molecules has only been made possible through the introduction of automation. In this respect we have developed a fully automatic microscope which is integrated with a robotic liquid handling station. Results from the first series of assays have already allowed us to identify proteins which localise to distinct organelles of the secretory pathway and appear to be new regulators of different steps in protein transport. Molecular interaction studies with these factors has extended our screens and identified novel and known interactors. Altogether, this approach will ultimately allow us to identify functional networks of proteins in a morphological context, and will greatly contribute to our understanding of whole cell function.  

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List of Abstracts * Speakers * Organizers * Authors * Original Announcement

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